MojoSort™ Nanobeads No Wash Protocol
Prepare cells from your tissue of interest without lysing erythrocytes.
In the final wash of your sample preparation, resuspend the cells in MojoSort™ Buffer by adding up to 4 mL in a 5 mL (12 x 75 mm) polystyrene tube.
Note: Keep MojoSort™ Buffer on ice throughout the procedure.
Filter the cells with a 70 μm cell strainer, centrifuge at 300 x g for 5 minutes, and resuspend in an appropriate volume of MojoSort™ Buffer.
Count and adjust the cell concentration to 1 x 108 cells/mL.
Aliquot 100 μL of cell suspension (107 cells) into a new tube.
Resuspend the beads by vortexing, maximum speed, 5 touches.
Add 10 μL of Nanobeads, mix well and incubate on ice for 15 minutes.Scale up the volume accordingly if separating more cells.
For example, add 100 μL for 1 x 108 cells.
When working with less than 107 cells, use indicated volumes for 107 cells.
Resuspend the cells in 3 mL of MojoSort™ Buffer.
Optional: Take an aliquot before placing the tube in the magnet to monitor purity and yield.
Place the tube in the magnet for 5 minutes.
Pour out the liquid.
Resuspend labeled cells in appropriate buffer.
Repeat steps 6 – 8 on the labeled fraction 2 more times, for a total of 3 magnetic separations.Optional: Take a small aliquot to monitor purity and yield.
If desired, pool the unlabeled fractions and process simultaneously with the positive labeled cells when assessing purity and yield.
